Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
2001
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
2008
Deviations:
no
Qualifier:
according to
Guideline:
other: JMAFF guidelines including the most recent partial revisions
Version / remarks:
2011
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: F-12676, PDC lot 20
- Expiration date of the lot/batch: 03 September 2013 (allocated by WIL Research Europe B.V., 1 year after receipt of the test substance)
- Purity test date: 99.96%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature in the dark

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
Adjustment was made for specific gravity of the vehicle. No correction was made for purity of the test substance. In order to obtain homogeneity, the test substance (formulations) were heated in a water bath with a maximum temperature 66ºC for a maximum of 15 minutes. The test substance (formulations) were allowed to cool down to a temperature of maximally 40ºC prior to dosing.

FORM AS APPLIED IN THE TEST (if different from that of starting material): White solid material dissolved in propylene glycol

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: Approximately 8-9 weeks old
- Weight at study initiation: 165.2 g (average)
- Fasting period before study: Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substance
- Housing: Group housing of 3 animals per cage in labeled Makrolon cages (MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cageenrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom)
- Diet (e.g. ad libitum): Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany)
- Water (e.g. ad libitum): Free access to tap water
- Acclimation period: Acclimatization period was at least 5 days before start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): 40-70%
- Air changes (per hr): Approximately 15 room air changes/hour
- Photoperiod (hrs dark / hrs light): 12:12 light:dark cycle

IN-LIFE DATES: From: 16 November 2012 To: 06 December 2012

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 2000 mg/kg (10 mL/kg) bw
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: Based on trial formulations performed at WIL Research Europe and on test substance data supplied by the sponsor

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

DOSAGE PREPARATION (if unusual): The formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. Adjustment was made for specific gravity of the vehicle. No correction was made for purity of the test substance. In order to obtain homogeneity, the test substance (formulations) were heated in a water bath with a maximum temperature 66ºC for a maximum of 15 minutes. The test substance (formulations) were allowed to cool down to a temperature of maximally 40ºC prior to dosing.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The toxicity of the test substance was assessed by stepwise treatment of groups of 3 females. The
first group was treated at a dose level of 2000 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
6 females (2 groups of 3 animals)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: Mortality/viability observed twice daily and body weights taken on days 1 (pre-administration), 8, and 15
- Necropsy of survivors performed: yes
- Other examinations performed: Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. The symptoms were graded according to fixed scales and the time of onset, degree and duration were recorded:
Maximum grade 4: grading slight (1) to very severe (4)
Maximum grade 3: grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1)

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Hunched/flat posture, uncoordinated movements, piloerection, lethargy and/or slow breathing were noted in the animals between Days 1 and 3.
Body weight:
The body weight gain shown by the animals over the study period was considered to be normal.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Based on the results of the test, the oral LD50 of the test article is greater than 2000 mg/kg bw in rats.
Executive summary:

The acute oral toxicity potential of the test article was evaluated in Wistar rats. The study was performed in compliance with OECD GLP (1997). The test method was based OECD 423 (2001). The test material was dissolved in propylene glycol to achieve the appropriate dose. Fasted female rats (n = 3) received a single 2000 mg/kg oral dose of the test article via gavage at a volume of 10 mL/kg. As all animals survived that dosing, another group of 3 female rats were dosed in a similar manner. Observations for mortality were made twice daily and body weights were collected on Days 1, 8, and 15. Observations for clinical signs of toxicity were made daily. Necropsy was performed on all animals at termination. All animals survived for the duration of the study. Hunched/flat posture (3 animals, and piloerection (4 animals) was observed through Day 2. Uncoordinated movements, lethargy and slow breathing were noted in all animals on the day of exposure at up to 4 hours postdose. The body weight gains were considered normal. No gross abnormalities were observed at necropsy.

Based on the results of the test, the oral LD50 of the test article is greater than 2000 mg/kg bw in rats.